生物技术通报2016，Vol.32Issue(7)：138-145,8.DOI:10.13560/j.cnki.biotech.bull.1985.2016.07.021
蓝藻橙色类胡萝卜素蛋白的基因克隆及在大肠杆菌中的异源表达和功能分析
Gene Cloning of the Orange Carotenoid Protein from Cyanobacteria, and Its Ectopic Expression and Functional Evaluation in Escherichia coli
摘要
Abstract
The aim is to verify the orange carotenoid protein(OCP)in cyanobacteria can scavenge singlet oxygen(1O2)independent of carotenoids in vivo. The cyanobacterial OCP gene was amplified by PCR,and then subjected to prokaryotic inducible expression and further evaluation of anti-1O2 activity in Escherichia coli by bacterial dot-plating test and growth curve assay. Results showed that the gene(SyOCP) of OCP was successfully amplified from the metagenome of the cyanobacteria water,with a coding region of 954 bp. Bioinformatics analysis indicated that SyOCP was from the major cyanobacteria species—Synechocystis sp. PCC 6803. By IPTG induction,the SyOCP gene was highly expressed in E. coli with soluble protein. In addition,the ectopic expression of SyOCP in recombinant E. coli resulted in the notable enhancement of it on the tolerance to the common 1O2-induced photosensitizer dye,methylene blue. The intrinsic anti-1O2 role of cyanobacterial OCP(without binding carotenoids)was in vivo verified in E. coli.关键词
蓝藻/橙色胡萝卜素蛋白/宏基因组/单线态氧/亚甲基蓝Key words
cyanobacteria/orange carotenoid protein/metagenome/singlet oxygen/methyl blue引用本文复制引用
刘畅,罗著,张梦如,杨玉梅,刘娴,龚明,邹竹荣..蓝藻橙色类胡萝卜素蛋白的基因克隆及在大肠杆菌中的异源表达和功能分析[J].生物技术通报,2016,32(7):138-145,8.基金项目
国家自然科学基金项目 （）
